High resolution mapping of fluoroquinolones in TB rabbit lesions reveals specific distribution in immune cell types

  1. Landry Blanc
  2. Isaac B Daudelin
  3. Brendan K Podell
  4. Pei-Yu Chen
  5. Matthew Zimmerman
  6. Amanda J Martinot
  7. Rada M Savic
  8. Brendan Prideaux
  9. Veronique Anne Dartois  Is a corresponding author
  1. Rutgers, The State University of New Jersey, United States
  2. Colorado State University, United States
  3. Harvard Medical School, United States
  4. University of California, San Francisco, United States

Abstract

Understanding the distribution patterns of antibiotics at the site of infection is paramount to selecting adequate drug regimens and developing new antibiotics. Tuberculosis (TB) lung lesions are made of various immune cell types, some of which harbor persistent forms of the pathogen, Mycobacterium tuberculosis. By combining high resolution MALDI MSI with histology staining and quantitative image analysis in rabbits with active TB, we have mapped the distribution of a fluoroquinolone at high resolution, and identified the immune-pathological factors driving its heterogeneous penetration within TB lesions, in relation to where bacteria reside. We find that macrophage content, distance from lesion border and extent of necrosis drive the uneven fluoroquinolone penetration. Preferential uptake in macrophages and foamy macrophages, where persistent bacilli reside, compared to other immune cells present in TB granulomas, was recapitulated in vitro using primary human cells. A nonlinear modeling approach was developed to help predict the observed drug behavior in TB lesions. This work constitutes a methodological advance for the co-localization of drugs and infectious agents at high spatial resolution in diseased tissues, which can be applied to other diseases with complex immunopathology.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 3, 4 and 5. Model codes are provided for the base model and full model.

Article and author information

Author details

  1. Landry Blanc

    Public Health Research Insitute, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Isaac B Daudelin

    Public Health Research Insitute, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Brendan K Podell

    Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Pei-Yu Chen

    Public Health Research Insitute, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Matthew Zimmerman

    Public Health Research Institute, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Amanda J Martinot

    Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Rada M Savic

    Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Brendan Prideaux

    Public Health Research Insitute, Rutgers, The State University of New Jersey, Newark, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Veronique Anne Dartois

    Public Health Research Insitute, Rutgers, The State University of New Jersey, Newark, United States
    For correspondence
    veronique.dartois@rutgers.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9470-5009

Funding

National Institutes of Health (U01-HL131072)

  • Veronique Anne Dartois

National Institutes of Health (R01-AI111967)

  • Veronique Anne Dartois

Bill and Melinda Gates Foundation (OPP1174780)

  • Veronique Anne Dartois

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal studies were performed in Biosafety Level 3 facilities and approved by the Institutional Animal Care and Use Committee (IACUC protocol number 16016) of the New Jersey Medical School, Rutgers University, Newark, NJ, under the guidelines and regulations of the National Institutes of Health.

Reviewing Editor

  1. Bavesh D Kana, University of the Witwatersrand, South Africa

Publication history

  1. Received: August 15, 2018
  2. Accepted: November 13, 2018
  3. Accepted Manuscript published: November 14, 2018 (version 1)
  4. Version of Record published: November 21, 2018 (version 2)

Copyright

© 2018, Blanc et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,505
    Page views
  • 285
    Downloads
  • 34
    Citations

Article citation count generated by polling the highest count across the following sources: Scopus, Crossref, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Landry Blanc
  2. Isaac B Daudelin
  3. Brendan K Podell
  4. Pei-Yu Chen
  5. Matthew Zimmerman
  6. Amanda J Martinot
  7. Rada M Savic
  8. Brendan Prideaux
  9. Veronique Anne Dartois
(2018)
High resolution mapping of fluoroquinolones in TB rabbit lesions reveals specific distribution in immune cell types
eLife 7:e41115.
https://doi.org/10.7554/eLife.41115

Further reading

    1. Immunology and Inflammation
    2. Microbiology and Infectious Disease
    Mary F Fontana, Erica Ollmann Saphire, Marion Pepper
    Research Article

    Naturally acquired immunity to malaria develops only after many years and repeated exposures, raising the question of whether Plasmodium parasites, the etiological agents of malaria, suppress the ability of dendritic cells (DCs) to activate optimal T cell responses. We demonstrated recently that B cells, rather than DCs, are the principal activators of CD4+ T cells in murine malaria. In the present study, we further investigated factors that might prevent DCs from priming Plasmodium-specific T helper cell responses. We found that DCs were significantly less efficient at taking up infected red blood cells (iRBCs) compared to soluble antigen, whereas B cells more readily bound iRBCs. To assess whether DCs retained the capacity to present soluble antigen during malaria, we measured responses to a heterologous protein immunization administered to naïve mice or mice infected with P. chabaudi. Antigen uptake, DC activation, and expansion of immunogen-specific T cells were intact in infected mice, indicating DCs remained functional. However, polarization of the immunogen-specific response was dramatically altered, with a near-complete loss of germinal center T follicular helper cells specific for the immunogen, accompanied by significant reductions in antigen-specific B cells and antibody. Our results indicate that DCs remain competent to activate T cells during Plasmodium infection, but that T cell polarization and humoral responses are severely disrupted. This study provides mechanistic insight into the development of both Plasmodium-specific and heterologous adaptive responses in hosts with malaria.

    1. Genetics and Genomics
    2. Microbiology and Infectious Disease
    Lillian C Lowrey, Leslie A Kent ... Peggy A Cotter
    Research Article

    Adaptation to fluctuating environmental conditions is difficult to achieve. Phase variation mechanisms can overcome this difficulty by altering genomic architecture in a subset of individuals, creating a phenotypically heterogeneous population with subpopulations optimized to persist when conditions change, or are encountered, suddenly. We have identified a phase variation system in Burkholderia thailandensis that generates a genotypically and phenotypically heterogeneous population. Genetic analyses revealed that RecA-mediated homologous recombination between a pair of insertion sequence (IS) 2-like elements duplicates a 208.6 kb region that contains 157 coding sequences. RecA-mediated homologous recombination also resolves merodiploids, and hence copy number of the region is varied and dynamic within populations. We showed that the presence of two or more copies of the region is advantageous for growth in a biofilm, and a single copy is advantageous during planktonic growth. While IS elements are well-known to contribute to evolution through gene inactivation, polar effects on downstream genes, and altering genomic architecture, we believe that this system represents a rare example of IS element-mediated evolution in which the IS elements provide homologous sequences for amplification of a chromosomal region that provides a selective advantage under specific growth conditions, thereby expanding the lifestyle repertoire of the species.